Vascular stabilization as a broad theraprutic platform for biodefense

许多A、B和C类优先病原体的感染激活了先天免疫系统，触发了 大量释放细胞因子和其他渗透性因素，破坏内皮屏障的稳定。这个 血管完整性的丧失会导致非心源性水肿、休克、多器官衰竭和死亡。我们建议 一个新的广泛的治疗平台，以减少发病率和死亡率基于我们确定的 内皮受体，Robo4。Robo4受体被其配体Sit蛋白激活后，保留了 内皮屏障的完整性，并干扰来自多个 血管内皮细胞的通透性因子包括肿瘤坏死因子-α、白介素2和凝血酶。在动物模型中， Robo4在各种损伤后稳定内皮细胞，包括机械损伤，缺血和 细胞因子风暴。我们最近已经证明，在两种病原体诱导的急性肺损伤的啮齿动物模型中 受伤，这种方法减少了浮肿、休克和死亡。最终，我们的目标是为 治疗各种A、B和C类优先病原体的感染。 目的1：观察Robo4多肽激动剂对病毒性出血热啮齿动物模型的疗效。我们 将使用病毒出血热模型来检查在这些非流感病毒中激活Robo4 病原体(阿拉伯病毒、出血热病毒)可降低死亡率。这一目标是由一个积极的 已经在细菌内毒素和H5N1动物模型中建立了疗效的合作努力。 目的2：优化Robo4多肽激动剂。我们将优化重组狭缝样肽，使其能够 通过系列突变激活Robo4信号通路。研究将集中在交付、稳定性和 为未来的GMP/GLP研究做准备所需的生产要求。 目的3：ROBO信号在内皮和上皮完整性中的作用。我们将确定Sit-Robo4是否 增强内皮细胞与细胞之间的相互作用，并试图了解其分子机制。我们将使用 这是一个研究Sit-Robo1信号在上皮-细胞-细胞接触中是否发挥类似作用的模型。 该项目符合RMRCE关于病毒治疗的综合研究重点，并在 五年期间将与RP3.1、3.2、2.3以及C、D和F核心直接互动。

Infection by many Category A, B, and C Priority Pathogens activates the innate immune system triggering an exuberant release of cytokines and other permeability factors that destabilize the endothelial barrier. The loss of vascular integrity results in non-cardiogenic edema, shock, multi-organ failure and death. We propose a new and broad therapeutic platform for reducing morbidity and mortality based on our identification of an endothelial receptor, Robo4. Activation of the Robo4 receptor by its ligand , Slit proteins, preserves the integrity of the endothelial barrier and interferes with the downstream signaling cascades from multiple permeability factors including TNF-alpha, interleukins, and thrombin in the endothelium. In animal models, Robo4 stabilizes the endothelium following diverse insults including mechanical injury, ischemia, and cytokine storm. We have recently demonstrated that in two rodent models of pathogen induced acute lung injury, this approach reduced edema, shock and death. Ultimately, our goal is to create a single platform for treating infections from a variety of Category A, B and C priority pathogens. AIM 1: Determine efficacy of Robo4 peptide agonist in rodent models of viral hemorrhagic fevers. We will use viral hemorrhagic fever models to examine whether activating Robo4 in these non-influenza viral pathogens (arenavirus hemorrhagic fever virus) reduces mortality. This Aim is facilitated by an active collaborative effort that has already established efficacy in bacterial endotoxin and H5N1 animal models. AIM 2: Optimize a Robo4 peptide agonist. We will optimize recombinant Slit-like peptide capable of activating the Robo4 signaling pathway by serial mutagenesis. Studies will focus on delivery, stability, and production requirements needed to prepare for future GMP/GLP studies. AIM 3: Robo signaling in endothelial and epithelial integrity. We will determine whether Slit-Robo4 enhances endothelial cell-cell interactions, and seek to understand the molecular mechanism. We will use this as a model to investigate whether Slit-Robo1 signaling plays a similar role in epithelial-cell-cell contacts. This project fits within the RMRCE Integrated Research Focus on Viral Therapeutics and over the course of the five year period will interact directly with RP3.1, 3.2, 2.3 and cores C, D, and F.